OBJECTIVE To identify the quality markers （Q-Markers） of Hyssopus cuspidatus against airway remodeling in bronchial asthma （referred to as “asthma”）， an d to provide a reference for the quality control research of H. cuspidatus based on pharmacodynamic activity. METHODS Potential active components and action targets of H. cuspidatus were screened by network pharmacology method. Using human airway smooth muscle cells （HASMCs） as objects， airway remodeling cell model was induced by platelet-derived growth factor-BB （PDGF-BB）； validation test was then performed for anti-asthmatic effects of H. cuspidatus and the potential active components. HPLC method was employed to establish the fingerprints of 14 batches of H. cuspidatus samples， and chemometric analysis was also conducted. Combined with the results of pharmacodynamic experiments and fingerprint analysis， the Q-Markers of H. cuspidatus against airway remodeling in asthma were determined. RESULTS&CONCLUSIONS Network pharmacology analysis showed that the potential active components of H. cuspidatus against asthma might be flavonoids and phenolic acids such as luteolin， quercetin and rosmarinic acid， and the core anti-asthmatic targets were interleukin-6， mitogen-activated protein kinase， etc. In vitro experimental results confirmed that 25， 50， 100 μg/mL of H. cuspidatus ， as well as neochlorogenic acid （80 μmol/L）， acacetin （80 μmol/L）， salvianolic acid B （40 μmol/L） and quercetin-3- O - β -D-glucuronide （80 μmol/L）， significantly reduced the cell viability induced by PDGF-BB， inhibited cell proliferation， migration， and the phosphorylation level of extracellular signal-regulated protein kinase 1/2， decreased the levels of interleukin-6， tumor necrosis factor-α and reactive oxygen species， and generally arrested cells in the G 0 /G 1 phase （ P ＜0.05）. Fingerprint analysis showed that there were 27 common peaks in the fingerprints of the 14 batches of H. cuspidatus samples， with 15 compounds （including luteolin） identified， and the similarities of fingerprints were all greater than 0.8. The 14 batches of samples could be divided into three categories： S1-S7 as one category， S8-S13 as one category， and S14 as one category. The variable importance in the projection values of rosmarinic acid， chlorogenic acid， caffeic acid， salvigenin， luteolin， ferulic acid， quercetin-3- O - β -D-glucuronide， and the components corresponding to peaks 5 and 8 were greater than 1， indicating they were potential differential markers affecting quality. Integrating network pharmacology， in vitro experimental validation， and chemometric analysis， rosmarinic acid， neochlorogenic acid， caffeic acid， salvigenin， luteolin， ferulic acid， quercetin-3- O - β -D-glucuronide， acacetin， salvianolic acid B and chlorogenic acid may be the Q-Markers of H. cuspidatus against asthma.

The disease-syndrome combination animal model in traditional Chinese medicine （TCM） andrology serves as an important bridge linking TCM theory with modern medical research， providing a key experimental platform for elucidating the 'syndrome-disease' correlation mechanism in male-specific diseases and for screening effective prescriptions. This article reviews recent progress in animal model research on common TCM andrological diseases， including prostatic diseases， sexual dysfunction， and male infertility， with a focus on analyzing the application， advantages， and disadvantages of various modeling strategies， such as immune induction， hormonal intervention， and multi-factor combination across different syndrome types. However， despite breakthroughs in model construction techniques， current research still faces several challenges， including insufficient standardization of syndrome differentiation and difficulties in quantifying TCM-specific indicators. Future studies need to optimize model evaluation systems by integrating modern technologies， in order to promote the standardization and internationalization of TCM andrology research.

To review the clinical characteristics, short-term outcomes, and risk factors of patients with infective endocarditis(IE) who underwent surgical treatment at a single center, and to summarize treatment experience.

Objective To evaluate the clinical effect of intensive treatment of obese patients with type 2 diabetes with Degu asparagus insulin and semaglutide.Methods A total of 92 obese patients with type 2 diabetes who were admitted to Rugao Hospital of Traditional Chinese Medicine from May 2020 to May 2023 were selected and randomly divided into two groups by randomnumber table method,with 46 cases in each group.The treatment group received Degu asparagus insulin and semaglutide,and the control group was treated with semaglutide.Glucose and lipid metabolism indicators(fasting blood glucose,glycated hemoglobin,fructosamine,2-hour postprandial blood glucose,and total cholesterol),blood glucose fluctuations(standard deviation of blood glucose,amplitude of postprandial blood glucose fluctuations,24-hour average blood glucose),insulin resistance index,visceral fat index,antioxidant indicators(malondialdehyde[MDA],lipid peroxide[LPO],superoxide dismutase[SOD],and paraoxonase-1[PON1]),and adverse reactions were observed before and after 4 weeks of treatment.Results After 4 weeks of treatment,the treatment group showed a significant improvement in glucose and lipid metabolism compared to the control group,including decreased fasting blood glucose,glycated hemoglobin,fructosamine,2-hour postprandial blood glucose,and total cholesterol(P<0.05).In addition,the treatment group showed significant reduction in the standard deviation of blood glucose,amplitude of postprandial blood glucose fluctuations,and 24-hour average blood glucose.Insulin resistance and visceral fat index were also significantly decreased in the treatment group(P<0.05).The decreases in MDA and LPO and the increases in SOD and PON1 indicated that the treatment group had better antioxidant capacity(P<0.001).There was no significant difference in the incidence of gastrointestinal adverse reactions,hypoglycemia,or liver damage between the two groups(P>0.05).Conclusion Degu asparagus insulin combined with semaglutide can effectively improve metabolic indicators of obese patients with type 2 diabetes and it provide an effective program for the comprehensive treatment of obese type 2 diabetes.

Objective To explore the therapeutic effect of LuoFuShan Rheumatism Plaster(LFS)on neuropathic pain(NP)and its molecular mechanism.Methods Mouse models of sciatic nerve chronic constriction injury(CCI)were treated with low,medium,and high doses(2.2,4.4,and 8.8 cm2,respectively)of LFS by topical application for 14 consecutive days.The therapeutic effects were assessed by evaluating the mechanical withdrawal threshold(MWT),paw withdrawal latency(PWL),plasma IL-6 and TNF-α levels,and histopathology of the sciatic nerve.Network pharmacology and molecular docking were used to identify the key targets and signaling pathways.The key targets were verified by RT-qPCR and immunohistochemistry.The biosafety of LFS was evaluated by measuring the organ indices and damage indicators of the heart,liver,and kidneys.Results Compared with the CCI group,LFS dose-dependently increased MWT and PWL,reduced plasma IL-6 and TNF-α levels,and alleviated sciatic nerve inflammation in the mouse models.Network pharmacology identified 378 bioactive compounds targeting 279 NP-associated genes enriched in TLR and TNF signaling.Molecular docking showed that quercetin and ursolic acid in LFS could stably bind to TLR4 and TNF-α.In the mouse models of sciatic nerve CCI,LFS significantly downregulated the mRNA expression levels of Tlr4 and Tnf-α in the spinal cord in a dose-dependent manner and lowered the protein expressions of TLR4 and TNF-α in the sciatic nerve.LFS treatment did not cause significant changes in the organ indices or damage indicators of the heart,liver and kidneys as compared with those in the CCI model group and sham-operated group.Conclusion LFS alleviates NP in mice by suppression of TLR4/TNF-α-mediated neuroinflammation with a good safety profile.

OBJECTIVES: To explore the therapeutic effect of LuoFuShan Rheumatism Plaster (LFS) on neuropathic pain (NP) and its molecular mechanism. METHODS: Mouse models of sciatic nerve chronic constriction injury (CCI) were treated with low, medium, and high doses (2.2, 4.4, and 8.8 cm², respectively) of LFS by topical application for 14 consecutive days. The therapeutic effects were assessed by evaluating the mechanical withdrawal threshold (MWT), paw withdrawal latency (PWL), plasma IL-6 and TNF-α levels, and histopathology of the sciatic nerve. Network pharmacology and molecular docking were used to identify the key targets and signaling pathways. The key targets were verified by RT-qPCR and immunohistochemistry. The biosafety of LFS was evaluated by measuring the organ indices and damage indicators of the heart, liver, and kidneys. RESULTS: Compared with the CCI group, LFS dose-dependently increased MWT and PWL, reduced plasma IL-6 and TNF-α levels, and alleviated sciatic nerve inflammation in the mouse models. Network pharmacology identified 378 bioactive compounds targeting 279 NP-associated genes enriched in TLR and TNF signaling. Molecular docking showed that quercetin and ursolic acid in LFS could stably bind to TLR4 and TNF‑α. In the mouse models of sciatic nerve CCI, LFS significantly downregulated the mRNA expression levels of Tlr4 and Tnf-α in the spinal cord in a dose-dependent manner and lowered the protein expressions of TLR4 and TNF-α in the sciatic nerve. LFS treatment did not cause significant changes in the organ indices or damage indicators of the heart, liver and kidneys as compared with those in the CCI model group and sham-operated group. CONCLUSIONS LFS alleviates NP in mice by suppression of TLR4/TNF-α-mediated neuroinflammation with a good safety profile.
Animals ; Toll-Like Receptor 4/metabolism* ; Neuralgia/metabolism* ; Mice ; Signal Transduction/drug effects* ; Tumor Necrosis Factor-alpha/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Sciatic Nerve/injuries* ; Male ; Molecular Docking Simulation ; Disease Models, Animal ; Interleukin-6

Objective The mouse autologous tumor model H22 is more valuable for tumor immunological-related research.This paper aims to establish mouse hepatic tumor cell line(H22-luc2-tdT)that stably express the tan-dem-dimer tomato(tdTomato)and luciferase genes.Establish an in vivo imaging model of cell line derived trans-planted tumors.Methods Using transplanted H22 tumor tissue,primary culture and continuous passage in vitro were performed to establish a continuous cell line.Cell proliferation,chromosome analysis,organoid culture,tumorigenicity,HE and ICH of aFP,CK7,CK15 were performed to charaterize the cell line.Then the luc2-tdT plasmid was transfected into H22 cells of P22,flow cytometry and in vitro/in vivo imaging were employed to screen and verify fluorescence expression.Mycoplasma detection and species verification of the established cell lines were performed.Results The H22 cells had been continuously passaged over 50 times.The cells of passsge 22(P22)were transplanted subcutaneously and intraperitoneally into C57 and Kunming mice,with a 100%tumor formation.The HE morphology of subcutaneous transplanted tumor were consistent with the original tumor.CK+/AFP+proved that it was of liver cancer origin.The H22 cells were hypo-triploid with a modal number of 40-44 chromosomes and telocentromeres,verifing its mouse origin.The latent phase for in vitro growth of H22 lasted from d0 to d3,while the exponential phaes d3 to d5,and reach plateou at d6.Successful transfection of H22 cells with the luc2-tdT were observed with in vitro/in vivo 100%fluorescence positivity,thus named H22-luc2-tdT.The transplanted tumor tissue of H22 cells could be primarily cultured to form organoids.The detection of Mycoplasma was negative,and its mouse origin confirmed by PCR.Conclusions H22 and H22-luc2-tdT cell lines are established and characterized,which can be used for the establishment and application of in vitro and in vivo liver cancer research and metastatic cell tracking.These cell lines are deposited at and can obtain from the National Biomedical Cell Resource Center(http://www.cellresource.cn).

Objective To establish primary and simian virus 40(SV40)T antigen transformed human vascular en-dothelial cell models,and provide available resources for endothelial research.Methods Human umbilical vein endothelial cells(HUVEC),human umbilical artery endothelial cells(HUAEC),great saphenous vein endothelial cells(GSVEC)and endothelial cells form endometrium and liver tissue were isolated and cultured respectively.Then,the primary endothelial cells were transformed by lentivirus containing SV40 big T and small T antigens,and continuously subcultured in vitro.The expression of CD31 was detected by flow cytometry,species identification-and mycoplasma detection by PCR,and cell identity was identified by STR detection.The transformed ECs were checked for HLA types.Some of them were tested for RNA expression profile and infected by Cas9 lentivirus to es-tablish stable clones.Results Totally 187 cell lines of transformed HUVEC,1 of transformed HUAEC,5 of trans-formed GSVEC,1 of transformed endothelial cells from endometrium and 1 of transformed endothelial cells from liv-er tissue,and 9 monoclonal HUVEC cell lines stably expressing Cas9 protein were established.All the transformed umbilical endothelial cells were CD31 positive ranging from 20%-90%for 20 cases,while for the rest 168 cases the positive rate was more than 90%.RNA expression revealed stable activation of cell proliferation(cell cycle and DNA synthesis).Their species were identified as human origin.The STR results were consistent with those of the primary culture and unique,and there was no mycoplasma contamination.All these cells could be obtained with the sharing services of National Science and Technology Infrastructure,the National Biomedical Cell-line Resource cen-ters(NSTI-BMCR).Conclusions A series of primary and SV40 T antigen transformed human vascular endothelial cell models have been established,which provide a tool for the study of cardiovascular diseases,inflammation,tumors and immune-related diseases.

Objective To compare the changes in anorectal function between patients with Parkinson's disease constipation(PDC)and with functional constipation(FC),and further understand the constipation characteristics of PDC patients.Methods From 2017 to 2022,34 patients with PDC and 39 patients with FC who visited the Department of Neurology and the Department of Gastroenterology in Xuanwu Hospital,Capital Medical University were selected as the observation group and the control group.High resolution anorectal manometry was used to detect the motility,sensation and reflex of anorectal sphincter and pelvic floor muscles in the two groups of patients with constipation,and to compare the anorectal function changes of the two groups of patients.according to the results of manometry,the characteristics of constipation in the two groups of patients were analyzed and compared.Results The resting anal sphincter pressure and intrarectal pressure in the PDC group were significantly lower than those in the FC group,with statistical significance[(71.73±20.01)mmHg vs(85.02±19.74)mmHg,(22.30±21.12)mmHg vs(38.10±17.00)mmHg,respectively,1 mmHg=0.133 kPa,all P＜0.05].The length of high pressure zone,maximum squeeze pressure,continuous systolic time and anal relaxation rate in PDC group were lower than those in FC group,but no statistically significant differences;The maximum tolerance threshold of patients in the FC group was significantly higher than that in the PDC group,but there was no statistically significant difference between the two groups.There was no statistically significant difference in rectanal inhibitory reflex,initial sensory threshold,initial defecation threshold,and rectal compliance between the two groups.According to the pressure measurement results,it was found that both groups of patients mainly had insufficient defecation thrust.The proportions of insufficient defecation propulsion in PDC group and FC group were 88.2％(30/34)and 59.0％(23/39),the proportions of dyscoordination were 11.8％(4/34)and 30.8％(12/39),and the proportions of normal defecation propulsion without uncoordinated defecation were 0％and 10.3％(4/39),respectively.There was significant difference between the two groups in the type composition ratio of defecation disorder(x2=8.623,P＜0.05).Conclusions Both PDC patients and FC patients have abnormal rectal and anal motility and sensation,and the main manifestation of pressure measurement classification is insufficient defecation thrust.However,compared to FC patients,PDC patients have significantly lower anal resting pressure and rectal defecation pressure.In depth research on the changes in anorectal function and constipation characteristics of PDC patients can help improve understanding of the disease and provide relevant evidence for developing reasonable treatment plans for PDC.

Objective To compare the changes in anorectal function between patients with Parkinson's disease constipation(PDC)and with functional constipation(FC),and further understand the constipation characteristics of PDC patients.Methods From 2017 to 2022,34 patients with PDC and 39 patients with FC who visited the Department of Neurology and the Department of Gastroenterology in Xuanwu Hospital,Capital Medical University were selected as the observation group and the control group.High resolution anorectal manometry was used to detect the motility,sensation and reflex of anorectal sphincter and pelvic floor muscles in the two groups of patients with constipation,and to compare the anorectal function changes of the two groups of patients.according to the results of manometry,the characteristics of constipation in the two groups of patients were analyzed and compared.Results The resting anal sphincter pressure and intrarectal pressure in the PDC group were significantly lower than those in the FC group,with statistical significance[(71.73±20.01)mmHg vs(85.02±19.74)mmHg,(22.30±21.12)mmHg vs(38.10±17.00)mmHg,respectively,1 mmHg=0.133 kPa,all P＜0.05].The length of high pressure zone,maximum squeeze pressure,continuous systolic time and anal relaxation rate in PDC group were lower than those in FC group,but no statistically significant differences;The maximum tolerance threshold of patients in the FC group was significantly higher than that in the PDC group,but there was no statistically significant difference between the two groups.There was no statistically significant difference in rectanal inhibitory reflex,initial sensory threshold,initial defecation threshold,and rectal compliance between the two groups.According to the pressure measurement results,it was found that both groups of patients mainly had insufficient defecation thrust.The proportions of insufficient defecation propulsion in PDC group and FC group were 88.2％(30/34)and 59.0％(23/39),the proportions of dyscoordination were 11.8％(4/34)and 30.8％(12/39),and the proportions of normal defecation propulsion without uncoordinated defecation were 0％and 10.3％(4/39),respectively.There was significant difference between the two groups in the type composition ratio of defecation disorder(x2=8.623,P＜0.05).Conclusions Both PDC patients and FC patients have abnormal rectal and anal motility and sensation,and the main manifestation of pressure measurement classification is insufficient defecation thrust.However,compared to FC patients,PDC patients have significantly lower anal resting pressure and rectal defecation pressure.In depth research on the changes in anorectal function and constipation characteristics of PDC patients can help improve understanding of the disease and provide relevant evidence for developing reasonable treatment plans for PDC.